Objective To explore the blood drug concentration monitoring of sustained-release valproate(DK)in children with epilepsy,focusing on the selection of sampling time and evaluation of the results.Methods Two hundred and seventy-one children taking DK and 155 children taking sodium valproate syrup(VPA Syr)were involved and their serum were taken when achieved steady state to determine the valproic acid level using fluorescence polarization immunoassay.They were divided into 4 groups,which were DK taken once daily group(DK qd group,126 children),DK taken once daily at night and sampled on morning group(DK qn group,26 children),DK taken every 12 h group(DK q12 h group,119 children),VPA Syr q12 h group(155 children).Determine the proportion of the blood drug concentration of each group below,ithin and above the therapeutic range for valproate(50-100 mg/L)were determined.The data were analyzed by t test.Results The Cmin of DK qd group were(73.09?19.91)mg/L,significantly lower from the serum concentration of DK qn and sampled on morning group [(94.94?25.44)mg/L](P0.05).Conclusions DK qn should sampled at night before the night dose.The Cmin of DK q12 h was higher according to the therapeutic range,it's favorable range still needs clinical practice.

Objective To construct humanized monoclonal antibodies against CD 20 and check their affinity to CD 20 antigen and their anti-tumor activity.Methods Based on the computer model , human IgG1 candidates closest to rituximab in crystal structure were selected in the Protein Data Bank ( PDB) .With the selected human IgG 1 candidates as the frame , we modified and transplanted the complementarity determining region ( CDR) of rituximab .First,the target gene fragments were obtained by overlapping PCR.Then, the sequences of the light chains(L) and the heavy chains(H) were inserted in-to the pcDNA3.3 and pOptiVEC vectors.Next, the constructed clones were transfected into 293F cells through transient transfection.After a large-scale cell culture, the mAb was purified by affinity chromatography rProtein A column.The puri-ty and expression level of the humanized antibodies was tested by sodium dodecyl sulfate ( SDS)-polyacrylamide gelelectro-phoresis(PAGE).The affinity of the humanized antibodies to CD20 was assessed with Fortebio assay.Finally, the anti-tumor activity of the constructed antibodies was detected by checking the tumor growth inhibition of the nude mice transplan-ted with tumor .Results Three humanized monoclonal antibodies against CD 20 were expressed and purified successfully . In reducing SDS-PAGE, the antibodies exhibited two bands of approximately 25 ×103 and 55 ×103 , respectively.The band size of the antibodies matched the expected value.Fortebio assay revealed that the humanized antibodies could bind to CD20 with high affinity (rituximab:6.48 ×10 -9mol/L, L4H7:1.91 ×10 -9mol/L, L5H5:7.35 ×10 -10mol/L,and L5H7:1.91 ×10 -9mol/L).The tumor growth inhibition experiment showed that the anti-tumor activity of L5H7 mAb was better than that of rituximab .Conclusion Three humanized monoclonal antibodies against CD 20 have been successfully construc-ted and expressed.L5H7 mAb possesses high affinity for CD20 and a good ability to kill tumor cells.

OBJECTIVETo observe the expression of cyclin D1 in human adnoid cystic carcinoma and its correlation with clinical characters.

METHODSThe expression of cycline D1 was evaluated with immunohistochemical method in forty-one cases of human adnoid cystic carcinoma, 15 cases of PA and 12 cases of normal salivary gland tissue.

RESULTSThe expression of cyclin D1 in normal tissue was negative, significantly different from PA and ACC (P < 0.05). The expression level of PA was significantly lower than ACC (P < 0.05). High expression of cyclin D1 was correlated with clinical stage and histological classification (P < 0.05), but not with sex, age, tumor site, recurrence, metastasis.

CONCLUSIONHigh expression of cyclin D1 promotes the formation and development of ACC.

Carcinoma ; Carcinoma, Adenoid Cystic ; Cyclin D1 ; Female ; Humans ; Middle Aged ; Salivary Gland Neoplasms

Objective To evaluate the effect of health education on controlling endemic fluorosis in Shaanxi Province between 2004 and 2006. Methods Hanbin, Pingli, Ziyang and Hanyin were selected for the investigation in 2004;in addition to the four areas, Zhengping, Shiquan and Langao were also for the investigation in 2006. Two schools were selected in each area, and 2 villages as the investigation spots. Fifty pupils in the fifth grade in each school, and 30 housewives between 25 - 50 years old in each village were chosen as the research subjects. Referring to the health education questionnaire in Technique Scheme of Endemic Disease Prevention Granted by Central Government of 2004, the degree of health education of endemic fluorine, arsenic poisoning in pupils and housewives were investigated. Results In 2004 and 2006, the average mark of pupils in school was 54.7 and 83.6, the pass rate was 57.5% (230/400) and 90.2% (629/697), respectively;the average mark of housewives was 59.7 and 83.9, the passing rate was 59.6% (143/240), 87.6%(338/386) respectively, indicating that the outcome was improved obviously in 2006 compared to that in 2004. Conclusions In the past three years, health knowledge of endemic arsenic and fluorosis has been improved among pupils and housewives in these areas.

Objective To investigate the geographic distribution panem of patients with anorectal atresia/stenosis in China.in order to provide clue for research on its etiology.Methods Data were collected from Chinese Birth Defects Monitoring Network(CBDMN),which was a hospital-based congenital malformations registry system.From 2001 to 2005,all fetuses with more than 28 weeks of gestation and neonates up to 7 days of age,were monitored.Two-dimensional graphic cluster method was used to divide monitoring stations into difierent classes with the incidence rates of anorectal atresia/stenosis.Results The overall incidence of anorectal atresia/stenosis was 3.17 per 10000 during 2001 to 2005.The incidence was higher in Eastern than that in Mid or Western paas of China and tbe difierence was statistically significant(z=2.50,3.69;P=0.012,＜0.001).The monitoring stations were grouped into 6 classes.Class I was with Helongjiang,Jilin and Liaoling;Class Ⅱ was with Fujian,Guangdong,Hainan,Guangxi,and South Hunan and Jiangxi;ClassⅢwas with Beijing,Tianjin,Hebei,Shandong,and Noah Jiangsu and Anhui;Class Ⅳ was with Zhejiang,Shanghai,South Anhui and Jiangsu,Noah Hunan and Jiangxi,Hubei,Henan,Shanxi and Inner Mongolia,Class V was with Ningxia,Gansu and Qinghai;and Class Ⅳ was with Shaanxi,Sichuan,Chongqing,Yunnan,Guizhou.Xinjiang and Tibet.Conclusion Our findings discovered the geographic distribution patterns of patients with anorectal atresia/stenosis in China.It is important to further analyze the relevant environmental factors attached to it so a beRer regional monitoring system for anorectal atresia/stenosis can be operated.

The c-Abl nonreceptor tyrosine kinase is activated in the cellular responses to genotoxic, oxidative and other forms of stress. Using tagged forms of c-Abl, the present studies demonstrate that c-Abl forms homodimers in cells. The results show that the c-Abl N-terminal regions interact with the corresponding C-terminal regions of both partners in the dimmer. Specifically, the c-Abl SH3 domain binds to a proline-rich motif at amino acids 958-982 in the c-Abl C-terminal region. Deletion of the proline-rich motif disrupts dimmer formation. These findings provide the first evidence that c-Abl forms homodimers and indicate that homodimerization can contribute to the regulation of c-Abl activity.
Humans ; Protein Multimerization ; Proto-Oncogene Proteins c-abl ; genetics ; metabolism ; src Homology Domains

To investigate the adjuvant effect of plasmid DNA encoding superantigen SEA (D227A) (pmSEA) on immune responses induced by HBV DNA vaccine containing HBV preS2 and S antigen in BABL/c (H-2d). BALB/c mice were immunized intramuscular injection with HBV DNA vaccine (pHBVS2S) mixed with or without pmSEA plasmid. Antibodies againat HBV PreS2 and S antigen in the sera were accessed by Anti-HBs ELISA, and the HBsAg specific cytotoxic T lymphocytes (CTLs) activity was determined by 5 Chromium Release Assay. The HBs peptide-specific IFN-gamma secreting T cells were detected by ELISPOT. Anti-HBs antibody titers and CTLs activity in mice immunized with pmSEA + pHBVS2S group were significant higher (P < 0.05) than pHBVS2S DNA vaccine group. The ratio of IgG1/IgG2a (0.282) was apparently different from the group immunized with peptide (10). Mice immunized with HBV DNA vaccine plus adjuvant produce higher titer of IgG1 and IgG2a antibodies against HBV S antigen 1.36 and 1.73 time higher than that without adjuvant respectively. HBs peptide--specific IFN-gamma secreting T cells increased 2 - 3 times by the pmSEA adjuvant, compared to DNA vaccine group. HBV DNA vaccine (pHBVS2S) induces humoral and cellular immuno-responses in BALB/c mice, and the responses could be significantly boasted by the plasmid encoding mSEA. Therefore the pmSEA was a potential adjuvant for DNA vaccines.
Adjuvants, Immunologic ; Animals ; Enterotoxins ; immunology ; Hepatitis B ; immunology ; prevention & control ; therapy ; Hepatitis B Antibodies ; blood ; Hepatitis B Vaccines ; immunology ; Interferon-gamma ; secretion ; Mice ; Mice, Inbred BALB C ; Staphylococcus aureus ; immunology ; Superantigens ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Vaccination ; Vaccines, DNA ; immunology

This study was purposed to analyse the immunophenotypic characteristics of chronic myelomonocytic leukemia (CMML), myelodysplastic syndromes (MDS) and acute monocytic leukemia (AML-M5b) by using multiparameter flow cytometry, and to explore its significance in diagnosis and differential diagnosis. The immunophenotypic characteristics of bone marrow samples from 14 CMML patients, 48 MDS patients, 46 AML-M5b patients and 18 normal persons were analyzed and compared by multiparametric flow cytometry. The results showed that the ratio of monocytes in CMML patients was obviously higher than that in MDS, AML-M5b patients and normal persons (P < 0.05), but there was no statistically significant difference between bone marrow samples of MDS and AML-M5b patients as well as normal persons. The ratio of blast cells in MDS patients was obviously higher than that in normal persons (P < 0.05), but did not show significant difference as compared with CMML patients. The ratio of mature granulocytes in AML-M5b patients was obviously lower than that in CMML and MDS patients as well as normal person bone marrow (P < 0.05). Certain differences of CD45/SSC characteristics in MDS, AML-M5b and CMML patients were found in comparison with normal persons. The abnormal expression of CD2, CD56, and CD14 tailing phenomenon were observed in CMML patients in comparison with bone marrow samples of MDS, AML-M5b and normal persons (P < 0.05). Lack and decrease of CD15 expression in MDS and CMML patients was significant different from AML-M5b and normal persons marrow, abnormal expression rate of CD15 in CMML patients was higher than that in MDS patients (P < 0.05), the CD13/CD11b/CD16 abnormal expression of granulocytes was seen in both CMML and MDS patients, but there was no statistically significant difference between them. Other antigens showed abnormality of varying degrees, but did not have any statistical significance. It is concluded that MDS, CMML and AML-M5b displayed a certain degree of similarity, and also possess their own immunophenotype characteristics. Comprehensive analysis of immunophenotype by multiparameter flow cytometry may be important for differential diagnosis among CMML, MDS and AML-M5b. High percentage of monocytes, abnormal coexpression of CD2, CD56 and CD14 tailing phenomenon, lack or decrease of CD15 as well as abnormal expression of CD13/CD11b/CD16 in granulocytes may play important roles in diagnosis of CMML.
Case-Control Studies ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; methods ; Leukemia, Monocytic, Acute ; diagnosis ; immunology ; Leukemia, Myelomonocytic, Chronic ; diagnosis ; immunology ; Myelodysplastic Syndromes ; diagnosis ; immunology

OBJECTIVETo detective the relationship between cyclooxygenase-2 (COX-2) and angiogenesis in oral squamous cell carcinoma (OSCC) and its clinical significance through observing the expression of COX-2 and determining microvessel density (MVD) in OSCC.

METHODSPV-9000 immunohistochemistry was used to determine the expression of COX-2 and CD34, which was used to determine MVD, in 76 OSCC tissues and 12 normal oral mucosa tissues.

RESULTSOverexpression of COX-2 was detected in OSCC, and was more intense compared with normal epithelium (P < 0.001). The high expression of COX-2 in OSCC was related to neck lymphnode metastasis, tumor size, TNM stage and histological grade (P <0.05). The MVD value in COX-2-positive group was much higher than that in COX-2-negative group (P < 0.01) and that in normal oral mucosa tissues (P < 0.01).

CONCLUSIONThe high expression of COX-2 in OSCC was significantly associated with MVD, neck lymphnode metastasis, tumor size, TNM stage and histological grade. COX-2 might be one of the important factors in the angiogenesis of OSCC.

Adult ; Carcinoma, Squamous Cell ; Cyclooxygenase 2 ; Female ; Humans ; Immunohistochemistry ; Lymphatic Metastasis ; Male ; Microvessels ; Middle Aged ; Mouth Mucosa ; Mouth Neoplasms ; Neovascularization, Pathologic

OBJECTIVESTo investigate the role of focal adhesion kinase (FAK) in the pathogenesis of cardiac hypertrophy induced by hypertension.

METHODSUsing immunofluorescent labeling, confocal microscopy and Western blotting, the expression and subcellular localization of FAK in the cardiac myocytes of left ventricle were determined in 2, 6, 12, and 18 month-old rats with spontaneously hypertensive heart failure (SHHF) along with age-matched control Wistar-Kyoto (WKY) rats.

RESULTSThere was no significant difference of FAK expression between 2 month-old SHHF and WKY rats (50.5+/-6.9 vs. 49.8+/-5.0, n=6, P>0.05). In contrast with the control groups, the expression of FAK significantly increased in 6, 12 and 18 month-old SHHF rats (130.6+/-3.0 vs. 47.3+/-1.3, 144.7+/-5.4 vs. 46.4+/-3.1, 141.4+/-9.8 vs. 48.5+/-2.2, each groups n=6, P<0.05) with FAK protein primarily cumulated in the intercalated disks and nuclei.

CONCLUSIONSFAK may play a role in the cell signaling transduction leading to cardiac hypertrophy, presumably through regulations of hypertrophic gene transcription and RNA processing.

Animals ; Focal Adhesion Kinase 1 ; metabolism ; Heart Ventricles ; pathology ; Hypertension ; complications ; Hypertrophy, Left Ventricular ; enzymology ; etiology ; Male ; Microscopy, Confocal ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Signal Transduction